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Characterization of expressed pigmented core light harvesting complex (LH 1) in a reaction center deficient mutant of Blastochloris viridis

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Abstract

The utility of photosynthetically defective mutants in the purple photosynthetic bacterium Blastochloris viridis (formerly Rhodopseudomonas viridis)was demonstrated with construction of a reaction-center deficient mutant, LH 1-H. This LH 1-H mutant has a photosynthetic apparatus in which most of the puf operon genes were deleted, resulting in an organism containing only the genes for the light harvesting polypeptides and the H subunit of the reaction center. This B. viridisstrain containing a truncation of the puf operon was characterized by gel electrophoresis, lipid-to-protein ratio analysis, optical spectroscopy, electron paramagnetic resonance and transmission electron microscopy. Optical and electron paramagnetic resonance spectroscopies revealed no photoactivity in this LH 1-H mutant consistent with the absence of intact reaction centers. Electron paramagnetic resonance evidence for assembled LH 1 complexes suggested that the interactions between light harvesting polypeptide complexes in membranes were largely unchanged despite the absence of their companion reaction center cores. The observed increase in the lipid-to-protein ratio was consistent with modified interactions between LH 1s, a view supported by transmission electron microscopy analysis of membrane fragments. The results show that B. viridis can serve as a practical system for investigating structure-function relationships in membranes and photosynthesis through the construction of photosynthetically defective mutants.

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  1. Agnes E. Ostafin

    Present address: Department of Chemical Engineering, University of Notre Dame, 182 Fitzpatrick, Notre Dame, IN, 46556, USA

Authors and Affiliations

  1. Department of Chemistry, Institute for Biophysical Dynamics, University of Chicago, 5735 S. Ellis Avenue, Chicago, IL, 60637, USA

    Nina S. Ponomarenko, Julia A. Popova, Martin Jäger & James R. Norris Jr.

  2. KAIROS Scientific, 10225 Barnes Canyon Road #A110, San Diego, CA, 92121, USA

    Edward J. Bylina

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  1. Agnes E. Ostafin
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  2. Nina S. Ponomarenko
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Ostafin, A.E., Ponomarenko, N.S., Popova, J.A. et al. Characterization of expressed pigmented core light harvesting complex (LH 1) in a reaction center deficient mutant of Blastochloris viridis . Photosynthesis Research 77, 53–68 (2003). https://doi.org/10.1023/A:1024921115267

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